skip to main content
DOE PAGES title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Xylose utilization stimulates mitochondrial production of isobutanol and 2-methyl-1-butanol in Saccharomyces cerevisiae

Abstract

Branched-chain higher alcohols (BCHAs), including isobutanol and 2-methyl-1-butanol, are promising advanced biofuels, superior to ethanol due to their higher energy density and better compatibility with existing gasoline infrastructure. Compartmentalizing the isobutanol biosynthetic pathway in yeast mitochondria is an effective way to produce BCHAs from glucose. However, to improve the sustainability of biofuel production, there is great interest in developing strains and processes to utilize lignocellulosic biomass, including its hemicellulose component, which is mostly composed of the pentose xylose. In this work, we rewired the xylose isomerase assimilation and mitochondrial isobutanol production pathways in the budding yeast Saccharomyces cerevisiae. We then increased the flux through these pathways by making gene deletions of BAT1, ALD6, and PHO13, to develop a strain (YZy197) that produces as much as 4 g/L of BCHAs (3.10 ± 0.18 g isobutanol/L and 0.91 ± 0.02 g 2-methyl-1-butanol/L) from xylose. This represents approximately a 28-fold improvement on the highest isobutanol titers obtained from xylose previously reported in yeast and the first report of 2-methyl-1-butanol produced from xylose. The yield of total BCHAs is 57.2 ± 5.2 mg/g xylose, corresponding to ~ 14% of the maximum theoretical yield. Respirometry experiments show that xylose increases mitochondrial activity by as muchmore » as 7.3-fold compared to glucose. The enhanced levels of mitochondrial BCHA production achieved, even without disrupting ethanol byproduct formation, arise mostly from xylose activation of mitochondrial activity and are correlated with slow rates of sugar consumption.« less

Authors:
 [1];  [2];  [1];  [1];  [1];  [1];  [2]; ORCiD logo [1]
  1. Princeton Univ., NJ (United States)
  2. Univ. of Illinois, Urbana-Champaign, IL (United States)
Publication Date:
Research Org.:
Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1571260
Grant/Contract Number:  
SC0018420
Resource Type:
Accepted Manuscript
Journal Name:
Biotechnology for Biofuels
Additional Journal Information:
Journal Volume: 12; Journal Issue: 1; Journal ID: ISSN 1754-6834
Publisher:
BioMed Central
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; isobutanol; xylose; 2-methyl-1-butanol; branched-chain higher alcohols; Saccharomyces cerevisiae; mitochondrial engineering

Citation Formats

Zhang, Yanfei, Lane, Stephan, Chen, Jhong-Min, Hammer, Sarah K., Luttinger, Jake, Yang, Lifeng, Jin, Yong-Su, and Avalos‬, José L. Xylose utilization stimulates mitochondrial production of isobutanol and 2-methyl-1-butanol in Saccharomyces cerevisiae. United States: N. p., 2019. Web. doi:10.1186/s13068-019-1560-2.
Zhang, Yanfei, Lane, Stephan, Chen, Jhong-Min, Hammer, Sarah K., Luttinger, Jake, Yang, Lifeng, Jin, Yong-Su, & Avalos‬, José L. Xylose utilization stimulates mitochondrial production of isobutanol and 2-methyl-1-butanol in Saccharomyces cerevisiae. United States. doi:10.1186/s13068-019-1560-2.
Zhang, Yanfei, Lane, Stephan, Chen, Jhong-Min, Hammer, Sarah K., Luttinger, Jake, Yang, Lifeng, Jin, Yong-Su, and Avalos‬, José L. Fri . "Xylose utilization stimulates mitochondrial production of isobutanol and 2-methyl-1-butanol in Saccharomyces cerevisiae". United States. doi:10.1186/s13068-019-1560-2. https://www.osti.gov/servlets/purl/1571260.
@article{osti_1571260,
title = {Xylose utilization stimulates mitochondrial production of isobutanol and 2-methyl-1-butanol in Saccharomyces cerevisiae},
author = {Zhang, Yanfei and Lane, Stephan and Chen, Jhong-Min and Hammer, Sarah K. and Luttinger, Jake and Yang, Lifeng and Jin, Yong-Su and Avalos‬, José L.},
abstractNote = {Branched-chain higher alcohols (BCHAs), including isobutanol and 2-methyl-1-butanol, are promising advanced biofuels, superior to ethanol due to their higher energy density and better compatibility with existing gasoline infrastructure. Compartmentalizing the isobutanol biosynthetic pathway in yeast mitochondria is an effective way to produce BCHAs from glucose. However, to improve the sustainability of biofuel production, there is great interest in developing strains and processes to utilize lignocellulosic biomass, including its hemicellulose component, which is mostly composed of the pentose xylose. In this work, we rewired the xylose isomerase assimilation and mitochondrial isobutanol production pathways in the budding yeast Saccharomyces cerevisiae. We then increased the flux through these pathways by making gene deletions of BAT1, ALD6, and PHO13, to develop a strain (YZy197) that produces as much as 4 g/L of BCHAs (3.10 ± 0.18 g isobutanol/L and 0.91 ± 0.02 g 2-methyl-1-butanol/L) from xylose. This represents approximately a 28-fold improvement on the highest isobutanol titers obtained from xylose previously reported in yeast and the first report of 2-methyl-1-butanol produced from xylose. The yield of total BCHAs is 57.2 ± 5.2 mg/g xylose, corresponding to ~ 14% of the maximum theoretical yield. Respirometry experiments show that xylose increases mitochondrial activity by as much as 7.3-fold compared to glucose. The enhanced levels of mitochondrial BCHA production achieved, even without disrupting ethanol byproduct formation, arise mostly from xylose activation of mitochondrial activity and are correlated with slow rates of sugar consumption.},
doi = {10.1186/s13068-019-1560-2},
journal = {Biotechnology for Biofuels},
number = 1,
volume = 12,
place = {United States},
year = {2019},
month = {9}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record

Save / Share:

Works referenced in this record:

Deconstruction of Lignocellulosic Biomass to Fuels and Chemicals
journal, July 2011


Saccharomyces cerevisiae Engineered for Xylose Metabolism Exhibits a Respiratory Response
journal, November 2004


Enzymatic assembly of DNA molecules up to several hundred kilobases
journal, April 2009

  • Gibson, Daniel G.; Young, Lei; Chuang, Ray-Yuan
  • Nature Methods, Vol. 6, Issue 5, p. 343-345
  • DOI: 10.1038/nmeth.1318

Non-fermentative pathways for synthesis of branched-chain higher alcohols as biofuels
journal, January 2008

  • Atsumi, Shota; Hanai, Taizo; Liao, James C.
  • Nature, Vol. 451, Issue 7174, p. 86-89
  • DOI: 10.1038/nature06450

Pyruvate decarboxylase: An indispensable enzyme for growth of Saccharomyces cerevisiae on glucose
journal, March 1996


Structure of the branched-chain keto acid decarboxylase (KdcA) from Lactococcus lactis provides insights into the structural basis for the chemoselective and enantioselective carboligation reaction
journal, November 2007

  • Berthold, Catrine L.; Gocke, Dörte; Wood, Martin D.
  • Acta Crystallographica Section D Biological Crystallography, Vol. 63, Issue 12, p. 1217-1224
  • DOI: 10.1107/S0907444907050433

Site-specific integration of DNA into wild-type and mutant lox sites placed in the plant genome
journal, April 1995


Features of promising technologies for pretreatment of lignocellulosic biomass
journal, April 2005


Branched-Chain Keto Acid Decarboxylase from Lactococcus lactis (KdcA), a Valuable Thiamine Diphosphate-Dependent Enzyme for Asymmetric C-C Bond Formation
journal, June 2007

  • Gocke, Dörte; Nguyen, Cong Luan; Pohl, Martina
  • Advanced Synthesis & Catalysis, Vol. 349, Issue 8-9, p. 1425-1435
  • DOI: 10.1002/adsc.200700057